Uncoordinated inhibition of gene expression for muscle proteins by a troponin T antisense oligodeoxynucleotide

The effect of antisense oligodeoxynucleotide to rat troponin T (TnT) mRNA on its expression in differentiated rat L6 myotubes in culture was examined. The target sequence following the initiation codon was between nucleotides 83 and 97 and is found in all mRNAs produced from the f-TNT gene. Our studies showed that chimeric oligomer with one phosphorothioate linkage at the 3'-end was considerably more resistant to nucleases than was a phosphodiester oligomer. The chimeric oligomer produced >50% inhibition of TnT polypeptide synthesis. Synthesis of myosin heavy chain (MHC), troponin I (TnI), and alpha and beta tropomyosins (Tm) was not inhibited by the anti-TnT oligomer. However, synthesis of alpha-actin and troponin C (TnC) was somewhat affected by this treatment. Furthermore, compared with the untreated control myotubes, the steady-state level of TnT mRNA was reduced by approximately 40%-50% in anti-TnT oligomer-treated myotubes. Cellular levels of three other muscle mRNAs, alpha-Tm, s-TnI, and alpha-actin were also reduced by approximately 30%-40%. In contrast, fast TnI, beta-Tm, and TnC mRNA levels were not significantly affected by this treatment. Therefore, inhibition of TnT synthesis in differentiated myotubes uncoupled the coordinated expression of muscle proteins.